Impact of co-culture on pancreatic differentiation of embryonic stem cells

Abstract

Promise of cellular therapy for type 1 diabetes has inspired the search for transplantable cell sources, and embryonic stem cells (ESCs) have emerged as strong candidates. We have developed a directed differentiation protocol to obtain insulin-producing cells from ESCs. The ESCs are first induced towards a homogeneous monolayer of definitive endoderm-like cells by co-culture with primary hepatocytes. Pancreatic commitment is induced by plating the ESC-derived endoderms on Matrigel, along with Sonic hedgehog inhibition and retinoid induction. More than 70% of differentiated cells positively upregulated Pdx-1, along with pro-endocrine transcription factors Ngn3, β2/neroD1, Nkx2.2 and Nkx6.1. Final maturation to islet-specific cells is achieved by co-culturing the ESC-derived pancreatic endocrine cells with endothelial cells, which resulted in Insulin 1 upregulation in 60% of the cell population, along with high levels of IAPP and Glut2. The differentiated cell population also secreted high levels of insulin. Our findings illustrate the significant effect of co-culture in different stages of differentiation and maturation of ESCs in vitro. Such a high yield of pancreatic islet cells has not yet been reported. Our findings establish a robust protocol for islet differentiation.

Long term effects of the implantation of autologous bone marrow mononuclear cells for type 2 diabetes mellitus

“…Given this relaies will be needed to substantiate these observations. Stem cells are a potential source of insulin-producing cells due to their capacity for proliferation and differentiation [29,30]. Bone marrow is an invaluable source of adult pluripotent stem cells, as it gives rise to hematopoietic stem cells, endothelial progenitor cells (EPCs), and mesenchymal stem cells (MSCs), amongst others.…”

Mind bomb 1 is required for pancreatic β-cell formation

“…Many protocols designed to induce development of insulin producing β-cells from human ES cells attempt to induce β-cell formation by inhibiting Notch activity with γ-secretase inhibitors (44)(45)(46)(47). Such protocols should now be reconsidered in light of our findings.…”

Endothelial Cell Co-culture Mediates Maturation of Human Embryonic Stem Cell to Pancreatic Insulin Producing Cells in a Directed Differentiation Approach

“…While the importance of endothelial cells during pancreatic development is increasingly being appreciated, its role in in vitro differentiation of embryonic cells is less investigated. In our earlier report we established the positive role of endothelial cells on pancreatic maturation of mouse embryonic stem cells 13 . In the current study we investigate the effect of endothelial cell signaling on differentiating human embryonic stem cells.…”